This revision application for supplemental funds is in response to Notice Number (NOT-OD-09-058) and Notice Title: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications. The parent INIA West U01 from which this application is derived is based on behavioral findings that the central amygdala nucleus (CeA) is a key brain area underlying stress reactions and the reinforcing properties of abused drugs, and that these behaviors involve several CeA transmitters (GABA, glutamate) and neuropeptides (CRF, opioids and galanin). We hypothesized that these neurochemical systems in the CeA are involved in the excessive ethanol drinking seen in dependent animals. Our electrophysiological studies of these systems in CeA neurons are either published (the role of CRF1, mu and delta opioid receptors) or ongoing (role of kappa opioid and galanin receptors). However, due to the recently stated need of the INIA West to begin physiological evaluation of the function of gene products suggested by the molecular components of the INIA West to be involved in excessive alcohol drinking, we propose the new studies of this application. Specifically, we will use electrophysiological methods to investigate the cellular role of the inflammatory factors identified by the Y. Blednov Research U01 (lipopolysaccharide {LPS}, TLR4, CD14, cytokines) in alcohol-related aspects of neuronal function in the CeA. Toward this aim we will use electrophysiological recording methods in brain slices in vitro and examine the interactions of LPS, ethanol and CRF on GABAergic and glutamatergic transmission in CeA neurons of wild type and CD14 knockout mice. These proposed new studies thus represent the new early steps toward evaluating the cellular sites and mechanisms of action of the emerging gene targets suggested by other INIA West components to underlie alcohol preference or excessive drinking. In addition, with this application we also are attempting to retain the job of our senior postdoctoral researcher and to allow full effort from our senior technician.